GSM7011567: NALM6_igg; Homo sapiens; OTHER
CUT and RUN data were generated using the Epicypher Cutana CUT&RUN kit v3.0 (14-1048) according to the manufacturers provided instructions. Briefly 500k NALM6 cells per reaction were bound to 10ml of provided activated ConA beads in 0.2ml PCR tubes. Bead-bound cells were suspended in Antibody Buffer (Wash buffer with 0.1% digitonin, 0.5mM Spermidine, 2mM EDTA, and 1x HALT protease inhibitors) and incubated with 1ul PU.1 antibody (Cell Signaling 2258) or IgG (Epicypher 13-0042k) overnight on a nutator mixer at 4C. The next day after washing, pAG MNase was bound and targeted digestion was carried out for 2 hours at 4C. Digestion was stopped using 33ml Stop buffer + 1ml (0.5ng) E.coli spike-in DNA and then cleaved DNA were released for 10 minutes at 37C. DNA were then purified for library preparation using the included purification kit. >30M 75bp paired end reads were generated per sample using the Illumina Novaseq.